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Production of MBP–HepA fusion protein in recombinant Escherichia coli by optimization of culture medium

Paper ID Volume ID Publish Year Pages File Format Full-Text
4905 256 2007 8 PDF Available
Title
Production of MBP–HepA fusion protein in recombinant Escherichia coli by optimization of culture medium
Abstract

Enhanced production of MBP (maltose-binding protein)-heparinase I (HepA) fusion protein in recombinant Escherichia coli was achieved by the optimization of the M9-based culture medium. First, by using the modified expression vector capable of releasing the C-terminus of HepA free to improve the specific activity of MBP–HepA, characteristics of the purified fusion protein were analyzed, which were similar to those of the native HepA except for the decreased affinity towards the substrate. M9-based culture medium was subsequently optimized for the enzyme production by orthogonal experimental design in shake flasks. Three major components were examined, namely glucose, yeast extract and calcium ion. The recombinant E. coli was further cultivated in a fermentor. As a result, total activity of HepA reached 20,650 IU l−1 in the optimized medium by a fed-batch mode in the 5-l fermentor. This study indicated that effective production of MBP–HepA by the present system would facilitate the large scale preparation of low molecular weight heparin (LMWH), which is a useful anticoagulant drug.

Keywords
Fusion protein; HepA; MBP; Orthogonal experimental design; Recombinant Escherichia coli; Vector modification
First Page Preview
Production of MBP–HepA fusion protein in recombinant Escherichia coli by optimization of culture medium
Publisher
Database: Elsevier - ScienceDirect
Journal: Biochemical Engineering Journal - Volume 34, Issue 2, May 2007, Pages 114–121
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering